Conjugate and use thereof

Abstract

A technique is provided which enables simple and low-cost purification of biologically active proteins by means of a conjugate of a target substance capturing molecule and a protein which comprises the amino acid sequence in SEQ ID NO:1, or a protein which comprises the amino acid sequence obtained by deleting, substituting or adding one or multiple amino acids in the amino acid sequence in SEQ ID NO: 1 and which has binding activity to a compound having —OH or —OR 1 [R 1 represents a hydrogen atom, an alkyl group or —PO 3 H 2 ].

1 . A conjugate of:
 a protein consisting of the amino acid sequence of SEQ ID NO: 1, or   a protein consisting of the amino acid sequence in which one or several amino acids have been deleted, substituted or added to the amino acid sequence of SEQ ID NO:   1, and having binding activity to a compound comprising —OH and —OR 1  wherein R 1  represents a hydrogen atom, an alkyl group, or —PO 3 H 2 ; and   a functional molecule.   
     
     
         2 . The conjugate of  claim 1 , wherein said functional molecule is a target substance capturing molecule. 
     
     
         3 . The conjugate of  claim 2 , wherein said target substance capturing molecule is an antibody, an antibody fragment, an aptamer, or a receptor against said target sub stance. 
     
     
         4 . The conjugate of  claim 2 , wherein said target substance capturing molecule is tumor necrosis factor receptor (TNFR). 
     
     
         5 . The conjugate of  claim 2 , wherein said target substance is a cytokine. 
     
     
         6 . A vector containing a nucleic acid encoding a fusion protein of:
 a protein consisting of the amino acid sequence of SEQ ID NO: 1, or   a protein consisting of the amino acid sequence in which one or several amino acids have been deleted, substituted or added to the amino acid sequence of SEQ ID NO: 1, and having binding activity to a compound comprising —OH and —OR 1  wherein R 1  represents a hydrogen atom, an alkyl group, or —PO 3 H 2 ; and   an antibody, an antibody fragment, a peptide aptamer, or a receptor against a target substance.   
     
     
         7 . A pharmaceutical composition comprising:
 a drug;   a carrier containing a compound comprising —OH and —OR 1  wherein R 1  represents a hydrogen atom, an alkyl group, or —PO 3 H 2 ; and   a conjugate of  claim 1  bound to said compound.   
     
     
         8 . A pharmaceutical containing a conjugate of  claim 1  as an active ingredient. 
     
     
         9 . A method of purifying a conjugate, the method comprising
 contacting a conjugate of  claim 1  with a compound comprising —OH and —OR 1  [R 1  represents a hydrogen atom, an alkyl group, or —PO 3 H 2 .] fixed to a solid phase.   
     
     
         10 . A solid phase for capturing a conjugate of  claim 1 , the solid phase characterized in that a compound comprising —OH and —OR 1 , wherein R 1  represents a hydrogen atom, an alkyl group, or —PO 3 H 2 , is fixed to said solid phase. 
     
     
         11 . A method of removing a conjugate of  claim 1 , the method comprising
 contacting plasma containing said conjugate with a compound comprising —OH and —OR 1 , wherein R 1  represents a hydrogen atom, an alkyl group, or —PO 3 H 2 , fixed to a solid phase.   
     
     
         12 . A method of removing a target substance, the method comprising contacting
 plasma containing a complex of a target substance and a conjugate of  claim 2  with   a compound comprising —OH and —OR 1 , wherein R 1  represents a hydrogen atom, an alkyl group, or —PO 3 H 2 , fixed to a solid phase.   
     
     
         13 . A method of removing a target substance, the method comprising contacting
 plasma containing a target substance with   a conjugate of  claim 2  fixed to a solid phase.   
     
     
         14 . A device for removing a conjugate of  claim 1 , the device comprising
 a removing means for removing said conjugate from plasma containing said conjugate, wherein said removing means comprises a solid phase to which a compound comprising —OH and —OR 1 , wherein R 1  represents a hydrogen atom, an alkyl group, or —PO 3 H 2 , is fixed.   
     
     
         15 . A device for removing a target substance, the device comprising
 a removing means for removing a complex of a target substance and a conjugate of  claim 2  from plasma containing said complex, wherein said removing means comprises a solid phase to which a compound comprising —OH and —OR 1 , wherein R 1  represents a hydrogen atom, an alkyl group, or —PO3H 2 , is fixed.   
     
     
         16 . A device for removing a target substance, the device comprising
 a removing means for removing a target substance from plasma containing said target substance, wherein said removing means comprises a solid phase to which a conjugate of  claim 2  is fixed.   
     
     
         17 . A labeling compound for detecting a conjugate of  claim 1 , the compound comprising —OH and —OR 1  wherein R 1  represents a hydrogen atom, an alkyl group, or —PO 3 H 2 . 
     
     
         18 . A method of detecting a conjugate, the method comprising
 forming a complex by allowing a labeling compound comprising —OH and —OR 1 , wherein R 1  represents a hydrogen atom, an alkyl group, or —PO 3 H 2 , to bind to a conjugate of  claim 1 ; and   detecting the labeling compound in said complex.   
     
     
         19 . A conjugate for detecting a compound comprising —OH and —OR 1 , wherein R 1  represents a hydrogen atom, an alkyl group, or —PO3H 2 , the conjugate characterized by being a conjugate of:
 a protein consisting of the amino acid sequence of SEQ ID NO: 1, or 
 a protein consisting of the amino acid sequence in which one or several amino acids have been deleted, substituted or added to the amino acid sequence of SEQ ID NO: 1, and having binding activity to a compound comprising —OH and —OR 1 , wherein R 1  represents a hydrogen atom, an alkyl group, or —PO 3 H 2 ; and 
 a labeling substance. 
 
     
     
         20 . A method of detecting a compound comprising —OH and —OR 1 , wherein R 1  represents a hydrogen atom, an alkyl group, or —PO 3 H 2 , the method comprising
 forming a complex by allowing a conjugate for detecting a compound of  claim 19  to bind to a compound comprising —OH and —OR 1 , wherein R 1  represents a hydrogen atom, an alkyl group, or —PO 3 H 2 ; and 
 detecting said labeling substance in said complex.